MicroED

Tamir Gonen on MicroED: In this interview, Tamir Gonen (UCLA) outlines the history of MicroED and its relationship to other methods for structure determination. While originally motivated by the desire to study membrane proteins, the up-and-coming method has now been applied to many different samples that often are not amenable to structural analysis by other means.

Cryo-electron microscopy (cryo-EM) methods promise new life to high-throughput macromolecular structure determination. Cryo-EM overcomes the fundamental barrier to X-ray diffraction determination of macromolecular complexes: growing X-ray grade crystals. Fortunately, the emergence of microcrystal electron diffraction (MicroED) facilitates the determination of new protein structures at atomic resolution from vanishingly small crystals.

Focusing the spotlight on MicroED: In this webinar, Tamir Gonen, Max Clabbers, and Emma Danelius provide insights into MicroED, discuss recent advances in method development, and present some new results.

MicroED exploits the strong interaction between electrons and nano-scale three-dimensional crystals and takes advantage of emerging cryo-EM instrumentation coupled to established crystallographic methods.

MicroED on a globular protein: proteinase K, a 30 kDa hydrolase which crystallizes in a tetragonal spacegroup, has been used extensively for methods development on MicroED.

Suggested further reading

2021

Martynowycz, Michael W; Clabbers, Max T B; Unge, Johan; Hattne, Johan; Gonen, Tamir

Benchmarking the ideal sample thickness in cryo-EM Journal Article

In: Proc Natl Acad Sci U S A, vol. 118, no. 49, 2021, ISSN: 1091-6490.

Abstract | Links | Altmetric | PlumX

2020

Zhu, Lan; Bu, Guanhong; Jing, Liang; Shi, Dan; Lee, Ming-Yue; Gonen, Tamir; Liu, Wei; Nannenga, Brent L

Structure Determination from Lipidic Cubic Phase Embedded Microcrystals by MicroED Journal Article

In: Structure, vol. 28, no. 10, pp. 1149–1159.e4, 2020, ISSN: 1878-4186.

Abstract | Links | Altmetric | PlumX

Zhu, Lan; Bu, Guanhong; Jing, Liang; Shi, Dan; Lee, Ming-Yue; Gonen, Tamir; Liu, Wei; Nannenga, Brent L

Structure Determination from Lipidic Cubic Phase Embedded Microcrystals by MicroED Journal Article

In: Structure, vol. 28, no. 10, pp. 1149–1159.e4, 2020, ISSN: 1878-4186.

Abstract | Links | Altmetric | PlumX

2018

Jones, Christopher G; Martynowycz, Michael W; Hattne, Johan; Fulton, Tyler J; Stoltz, Brian M; Rodriguez, Jose A; Nelson, Hosea M; Gonen, Tamir

The CryoEM Method MicroED as a Powerful Tool for Small Molecule Structure Determination Journal Article

In: ACS Cent Sci, vol. 4, no. 11, pp. 1587–1592, 2018, ISSN: 2374-7943.

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2013

Shi, Dan; Nannenga, Brent L; Iadanza, Matthew G; Gonen, Tamir

Three-dimensional electron crystallography of protein microcrystals Journal Article

In: Elife, vol. 2, pp. e01345, 2013, ISSN: 2050-084X.

Abstract | Links | Altmetric | PlumX

Suggested reviews

2021

Danelius, Emma; Gonen, Tamir

Protein and Small Molecule Structure Determination by the Cryo-EM Method MicroED Journal Article

In: Methods Mol Biol, vol. 2305, pp. 323–342, 2021, ISSN: 1940-6029.

Abstract | Links | Altmetric | PlumX

Martynowycz, Michael W; Gonen, Tamir

Studying Membrane Protein Structures by MicroED Journal Article

In: Methods Mol Biol, vol. 2302, pp. 137–151, 2021, ISSN: 1940-6029.

Abstract | Links | Altmetric | PlumX

Mu, Xuelang; Gillman, Cody; Nguyen, Chi; Gonen, Tamir

An Overview of Microcrystal Electron Diffraction (MicroED) Journal Article

In: Annu Rev Biochem, vol. 90, pp. 431–450, 2021, ISSN: 1545-4509.

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2017

Liu, Shian; Hattne, Johan; Reyes, Francis E; Sanchez-Martinez, Silvia; de la Cruz, M Jason; Shi, Dan; Gonen, Tamir

Atomic resolution structure determination by the cryo-EM method MicroED Journal Article

In: Protein Sci, vol. 26, no. 1, pp. 8–15, 2017, ISSN: 1469-896X.

Abstract | Links | Altmetric | PlumX

2016

Nannenga, Brent L; Gonen, Tamir

MicroED opens a new era for biological structure determination Journal Article

In: Curr Opin Struct Biol, vol. 40, pp. 128–135, 2016, ISSN: 1879-033X.

Abstract | Links | Altmetric | PlumX

2014

Nannenga, Brent L; Shi, Dan; Leslie, Andrew G W; Gonen, Tamir

High-resolution structure determination by continuous-rotation data collection in MicroED Journal Article

In: Nat Methods, vol. 11, no. 9, pp. 927–930, 2014, ISSN: 1548-7105.

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Protocols

2021

Martynowycz, Michael W; Gonen, Tamir

Protocol for the use of focused ion-beam milling to prepare crystalline lamellae for microcrystal electron diffraction (MicroED) Journal Article

In: STAR Protoc, vol. 2, no. 3, pp. 100686, 2021, ISSN: 2666-1667.

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2017

de la Cruz, M Jason; Hattne, Johan; Shi, Dan; Seidler, Paul; Rodriguez, Jose; Reyes, Francis E; Sawaya, Michael R; Cascio, Duilio; Weiss, Simon C; Kim, Sun Kyung; Hinck, Cynthia S; Hinck, Andrew P; Calero, Guillermo; Eisenberg, David; Gonen, Tamir

Atomic-resolution structures from fragmented protein crystals with the cryoEM method MicroED Journal Article

In: Nat Methods, vol. 14, no. 4, pp. 399–402, 2017, ISSN: 1548-7105.

Abstract | Links | Altmetric | PlumX

2016

Shi, Dan; Nannenga, Brent L; de la Cruz, M Jason; Liu, Jinyang; Sawtelle, Steven; Calero, Guillermo; Reyes, Francis E; Hattne, Johan; Gonen, Tamir

The collection of MicroED data for macromolecular crystallography Journal Article

In: Nat Protoc, vol. 11, no. 5, pp. 895–904, 2016, ISSN: 1750-2799.

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